Registration Dossier

Administrative data

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

Ames assay

Test chemical was evaluated for its mutagenic potential in Salmonella typhimurium TA100, TA1535, TA98 and TA1537 by AMES test. The test result was considered to be positive in all strain in the presence and absence of metabolic activation S9.

In vitro chromosomal abbreviation study

Test chemical was evaluated for its mutagenic potential in Chinese hamster ovary cells by in vitro mammalian chromosome aberration test. The test result was considered to be mutagenic both in the presence and absence of metabolic activation.

In vitro Mammalian cell gene mutation assay

Test chemical was evaluated for its mutagenic potential in mouse lymphoma by in vitro mammalian cell gene mutation. The test result was considered to be positive in Mouse lymphoma.

In vitro DNA damage and/or repair study

Test chemical was evaluated for its mutagenic potential in Chinese hamster ovary cells in vitro sister chromatid exchange assay. The test result was considered to be mutagenic both in the presence and absence of metabolic activation.

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Link to relevant study records

Referenceopen allclose all

Endpoint:
in vitro gene mutation study in bacteria
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
data from handbook or collection of data
Justification for type of information:
data from handbook or collection of data
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Principles of method if other than guideline:
To evaluate the mutagenic potential of test chemical in Salmonella Typhimurium TA98,TA 100,TA 1535 and TA 1537 by AMES test.
GLP compliance:
not specified
Type of assay:
bacterial reverse mutation assay
Target gene:
Histidine
Species / strain / cell type:
S. typhimurium, other: TA98,TA 100,TA 1535 and TA 1537
Details on mammalian cell type (if applicable):
not specified
Additional strain / cell type characteristics:
not specified
Cytokinesis block (if used):
not specified
Metabolic activation:
with and without
Metabolic activation system:
RLI = induced male Sprague Dawley rat liver S9 HLI = induced male Syrian hamster liver S9
Test concentrations with justification for top dose:
0,10,30,100, 333,1000 and 3333µg/plate
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: 95% Ethanol
- Justification for choice of solvent/vehicle: The test substance is soluble in 95% Ethanol.
Untreated negative controls:
not specified
Negative solvent / vehicle controls:
yes
Remarks:
95% Ethanol
True negative controls:
not specified
Positive controls:
yes
Positive control substance:
other: -S9 mix;4 nitro-O phenylenediamine (TA 98), Sodium azide (TA1535and TA 100) and 9-Aminoacridine(TA1537) +S9 mix; 2-Aminoanthracene(all strains)
Details on test system and experimental conditions:
Details on test system and conditions
METHOD OF APPLICATION: Pre incubation method
NUMBER OF REPLICATIONS: Duplicate
Evaluation criteria:
The plates were observed for revertents mutants colonies per plates
Statistics:
Yes, SD ± Mean was observed.
Key result
Species / strain:
S. typhimurium, other: TA100, TA1535, TA98 and TA1537
Metabolic activation:
with and without
Genotoxicity:
positive
Cytotoxicity / choice of top concentrations:
not specified
Vehicle controls validity:
valid
Untreated negative controls validity:
not specified
Positive controls validity:
valid
Remarks on result:
other: Mutagenic effect were observed

Strain: TA100

 

Dose

No Activation 
(Negative)

No Activation 
(Negative)

10% RLI 
(Negative)

10% RLI 
(Weakly Positive)

10% HLI 
(Positive)

10% HLI 
(Weakly Positive)

Protocol

Preincubation

Preincubation

Preincubation

Preincubation

Preincubation

Preincubation

ug/Plate

Mean

±SEM

Mean

±SEM

Mean

±SEM

Mean

±SEM

Mean

±SEM

Mean

±SEM

 0         

139

7.8

107

4

142

4.3

112

7.9

129

6.9

163

8.8

10         

121

5.4

146

3.2

33         

135

9.5

114

4.7

133

4.9

162

7.7

164

10.3

156

8.4

100         

145

8.4

117

11.8

132

14.2

169

10.2

210

8.7

223

11

333         

142

6.2

101

7.4

134

10

191

6.4

268

2.9

247

9

1000         

146

10.3

97

11.6

122

2.7

180

3.5

289

17.3

282

5.9

3333         

86 s

49.7

t

0 s

0

0 s

0

Positive Control

434

3.7

407

18.7

690

15.4

551

3.5

1290

45.3

1796

23.7

 

Strain: TA1535

Dose

No Activation 
(Negative)

10% RLI 
(Negative)

10% HLI 
(Negative)

Protocol

Preincubation

Preincubation

Preincubation

ug/Plate

Mean

±SEM

Mean

±SEM

Mean

±SEM

 0         

27

.9

46

3.8

42

4.5

33         

29

3.7

28

1.7

41

2.1

100         

25

3.5

21

1.8

36

.3

333         

27

3

16

1.2

37

2.2

1000         

28

5.2

0 s

0

32

3.5

3333         

9 s

3.1

t

0 s

0

Positive Control

427

7.1

277

7.1

536

12.8

 

Strain: TA1537

Dose

No Activation 
(Negative)

10% RLI 
(Negative)

10% HLI 
(Negative)

Protocol

Preincubation

Preincubation

Preincubation

ug/Plate

Mean

±SEM

Mean

±SEM

Mean

±SEM

 0         

5

.9

10

1

5

.6

33         

6

1

7

1.2

12

2.8

100         

6

1.7

7

.9

12

4

333         

6

.3

11

1.2

15

1.3

1000         

10

2

12

2.3

10

2

3333         

5

.3

9

1.7

0 s

0

Positive Control

172

4.7

167

14.8

500

5.5

 

Strain: TA98

Dose

No Activation 
(Negative)

No Activation 
(Positive)

10% RLI 
(Negative)

10% RLI 
(Positive)

10% HLI 
(Positive)

10% HLI 
(Positive)

Protocol

Preincubation

Preincubation

Preincubation

Preincubation

Preincubation

Preincubation

ug/Plate

Mean

±SEM

Mean

±SEM

Mean

±SEM

Mean

±SEM

Mean

±SEM

Mean

±SEM

 0         

21

2

29

2.2

40

1.7

37

3.5

37

3.7

37

4.3

10         

27

1.5

44

1.3

33         

26

3.1

52

7.2

38

4.2

53

.7

50

3

54

3.2

100         

30

2.7

64

4.6

50

2.3

57

1.9

67

3.6

62

3.7

333         

17

2.3

64

6.6

55

3.5

75

6.8

94

4.7

86

.3

1000         

17

1

84

7.1

56

4.7

83

1.2

242

19.6

113

13.5

3333         

t

0 s

0

t

0 s

0

Positive Control

724

34

724

7.1

425

15.3

382

27

1514

107.5

1027

53.1

 

Abbreviations:
RLI = induced male Sprague Dawley rat liver S9
HLI = induced male Syrian hamster liver S9
s = Slight Toxicity; p = Precipitate; x = Slight Toxicity and Precipitate; t = Toxic;

Conclusions:
Test chemical was evaluated for its mutagenic potential in Salmonella typhimurium TA100, TA1535, TA98 and TA1537 by AMES test. The test result was considered to be positive in all strain in the presence and absence of metabolic activation S9.
Executive summary:

Genetic toxicity in vitro study was assessed for test chemical. For this purpose AMES test was performed .The test material was exposed to Salmonella typhimurium TA100, TA1535, TA98 and TA1537in the presence and absence of metabolic activation S9. The concentration of test material used in the presence and absence of metabolic activation were 0, 10, 30,100, 333, 1000 and 3333 µg/plate. Mutagenic effects were observed in all strains, in the presence and absence of metabolic activation. Therefore test chemical was considered to be mutagenic in Salmonella typhimurium TA100, TA1535, TA98 and TA1537by AMES test. Hence the substance can be classified as gene mutant in vitro.

Endpoint:
in vitro cytogenicity / chromosome aberration study in mammalian cells
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
data from handbook or collection of data
Justification for type of information:
data from handbook or collection of data.
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)
Principles of method if other than guideline:
To evaluate the mutagenic potential of test chemical in Chinese hamster ovary cells by in vitro mammalian chromosome aberration test.
GLP compliance:
not specified
Type of assay:
other: in vitro mammalian chromosome aberration test
Target gene:
Not specified
Species / strain / cell type:
Chinese hamster Ovary (CHO)
Details on mammalian cell type (if applicable):
not specified
Additional strain / cell type characteristics:
not specified
Cytokinesis block (if used):
not specified
Metabolic activation:
with and without
Metabolic activation system:
induced male Sprague Dawley rat liver S9
Test concentrations with justification for top dose:
-S9;0,150,199,249and 300 µg/mL
+S9; 0,2460,2760,2990 and 3520 µg/mL
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: Dimethyl Sulfoxide
- Justification for choice of solvent/vehicle: The test substance is soluble in DMSO.
Untreated negative controls:
not specified
Negative solvent / vehicle controls:
yes
Remarks:
DMSO
True negative controls:
not specified
Positive controls:
yes
Positive control substance:
other: -S9 mix; Mitomycin-C +S9 mix; Cyclophosphamide
Details on test system and experimental conditions:
METHOD OF APPLICATION: In medium
DURATION
- Fixation time (start of exposure up to fixation or harvest of cells): 20.8 hours
NUMBER OF CELLS EVALUATED: 100 cells
Evaluation criteria:
The mammalian cells were observed for chromosome aberration, Chromosome gaps and breaks.
Statistics:
Yes, SD ± Mean was observed.
Key result
Species / strain:
Chinese hamster Ovary (CHO)
Metabolic activation:
with and without
Genotoxicity:
positive
Cytotoxicity / choice of top concentrations:
not specified
Vehicle controls validity:
valid
Untreated negative controls validity:
not specified
Positive controls validity:
valid
Remarks on result:
other: Mutagenic effects were observed


Dose
µg/mL

Total
Cells Examined

Total Aberrations

Complex Aberrations

Simple Aberrations

Other Abs

No.of
Abs

Abs
Per
Cell

% Cells
With
Abs

No.of
Abs.

Abs
Per
Cell

% Cells
With
Abs

No.of
Abs.

Abs
Per
Cell

% Cells
With
Abs

No.of
Abs.

Abs
Per
Cell

% Cells
With
Abs

Vehicle Control:

Negative (Not Specified)

0         

100

2

0.020

2.0

2

0.020

2.0

0

0.000

0.0

0

0.000

0.0

Dimethyl Sulfoxide

0         

100

2

0.020

2.0

2

0.020

2.0

0

0.000

0.0

0

0.000

0.0

Test Chemical:

2-Amino-4-nitrophenol

150         

100

12

0.120

3.0

0

0.000

0.0

1

0.010

1.0

11

0.110

2.0

199         

100

7

0.070

6.0

4

0.040

4.0

2

0.020

2.0

1

0.010

1.0

249         

100

12

0.120

11.0

6

0.060

6.0

3

0.030

3.0

3

0.030

3.0

300         

0

0

0.000

0.0

0

0.000

0.0

0

0.000

0.0

0

0.000

0.0

Positive Control:

Mitomycin-C

 0.0625    

50

18

0.360

26.0

16

0.320

24.0

2

0.040

4.0

0

0.000

0.0

Trend:

2.934

2.134

1.838

 

Probability:

0.002

0.016

0.033

 



Trial #:1_S9   Activation: Induced Rat Liver S9    Date: 03/10/1983    Harvest Time: 20.8 hour(s)   Trial Call: Positive   

 

Dose
µg/mL

Total
Cells Examined

Total Aberrations

Complex Aberrations

Simple Aberrations

Other Abs

 

No.of
Abs

Abs
Per
Cell

% Cells
With
Abs

No.of
Abs.

Abs
Per
Cell

% Cells
With
Abs

No.of
Abs.

Abs
Per
Cell

% Cells
With
Abs

No.of
Abs.

Abs
Per
Cell

% Cells
With
Abs

 

Vehicle Control:

Negative (Not Specified)

0         

100

2

0.020

2.0

1

0.010

1.0

0

0.000

0.0

1

0.010

1.0

 

Dimethyl Sulfoxide

0         

100

1

0.010

1.0

0

0.000

0.0

0

0.000

0.0

1

0.010

1.0

 

Test Chemical:

2-Amino-4-nitrophenol

2460         

100

14

0.140

5.0

1

0.010

1.0

1

0.010

1.0

12

0.120

3.0

 

2760         

100

103

1.030

28.0

35

0.350

19.0

15

0.150

11.0

53

0.530

8.0

 

2990         

50

74

1.480

34.0

16

0.320

18.0

6

0.120

10.0

52

1.040

14.0

 

3520         

0

0

0.000

0.0

0

0.000

0.0

0

0.000

0.0

0

0.000

0.0

 

Positive Control:

Cyclophosphamide

10         

50

10

0.200

18.0

6

0.120

12.0

4

0.080

8.0

0

0.000

0.0

 

Trend:

6.835

5.479

3.964

 

 

Probability:

0.000

0.000

0.000

 

 

Conclusions:
Test chemical was evaluated for its mutagenic potential in Chinese hamster ovary cells by in vitro mammalian chromosome aberration test. The test result was considered to be mutagenic both in the presence and absence of metabolic activation.
Executive summary:

Genetic toxicity in vitro study was assessed for test chemical. For this purpose in vitro mammalian chromosome aberration test was performed .The test material was exposed toChinese hamster ovary cells inthe presence and absence of metabolic activation S9. The concentration of test material used in the presence and absence of metabolic activation were mention below

-S9;0,150,199,249and 300 µg/mL

+S9; 0,2460,2760,2990 and 3520 µg/mL

Chromosome aberration, Chromosome gaps and breaks were observed in the presence or absence of metabolic activation. Therefore test chemical was considered to be mutagenic inChinese hamster ovary cells byin vitro mammalian chromosome aberration test. Hence the substance cannot be classified as mutagenic in vitro.

Endpoint:
in vitro gene mutation study in mammalian cells
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
data from handbook or collection of data
Justification for type of information:
data from handbook or collection of data
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 476 (In Vitro Mammalian Cell Gene Mutation Test)
Principles of method if other than guideline:
To evaluate the mutagenic potential of test chemical in mouse lymphoma by in vitro mammalian cell gene mutation.
GLP compliance:
not specified
Type of assay:
other: in vitro mammalian cell gene mutation test
Target gene:
Not specified
Species / strain / cell type:
other: mouse lymphoma cells
Details on mammalian cell type (if applicable):
not specified
Additional strain / cell type characteristics:
not specified
Cytokinesis block (if used):
not specified
Metabolic activation:
not specified
Test concentrations with justification for top dose:
Experiment I; 0,25,50,100,150,200 and 300 µg/mL
Experiment II: 50,100,150,200 and 300 µg/mL
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: Dimethyl Sulfoxide
- Justification for choice of solvent/vehicle: The test substance is soluble in DMSO.
Untreated negative controls:
not specified
Negative solvent / vehicle controls:
yes
Remarks:
DMSO
True negative controls:
not specified
Positive controls:
yes
Positive control substance:
other: Methyl Methane Sulfonate
Details on test system and experimental conditions:
METHOD OF APPLICATION: In culture dish
DETERMINATION OF CYTOTOXICITY; relative total growth
Evaluation criteria:
The mammalian cells were observed for mutagenic frequency in cells.
Statistics:
Yes, SD ± Mean was observed.
Key result
Species / strain:
other: mouse lymphoma cells
Metabolic activation:
not specified
Genotoxicity:
positive
Cytotoxicity / choice of top concentrations:
not specified
Vehicle controls validity:
valid
Untreated negative controls validity:
not specified
Positive controls validity:
valid
Remarks on result:
other: Mutagenic effects were observed

Nonactivation Trial: 1 Experiment Call: Positive

 

Conc.
ug/mL

Cloning
Efficiency

Relative Total
Growth

Mutant Colonies

Mutant Frequency

Avg Mutant Frequency

Vehicle Control

Dimethyl Sulfoxide

 0         

111

101

122

37

43

 

 

90

103

144

53

 

 

75

101

94

42

 

 

110

94

126

38

Test Chemical

2-Amino-4-nitrophenol

25         

70

53

198

94

98*

 

 

61

53

209

114

 

 

52

62

134

85

 

50         

72

53

233

109

108*

 

 

64

54

193

100

 

 

61

50

211

115

 

100         

65

27

349

179

194*

 

 

54

20

320

198

 

 

48

20

296

206

 

150         

63

4

501

266

271*

 

 

47

4

422

299

 

 

50

10

370

248

 

200         

273*

 

 

46

4

366

267

 

 

52

3

433

278

 

300         

 

 

 

 

Positive Control

Methyl Methane Sulfonate

5         

65

60

508

260

258*

 

 

77

26

598

258

 

 

65

60

503

257

 

Nonactivation Trial: 2 Experiment Call: Positive

 

Conc.
ug/mL

Cloning
Efficiency

Relative Total
Growth

Mutant Colonies

Mutant Frequency

Avg Mutant Frequency

Vehicle Control

Dimethyl Sulfoxide

 0         

93

90

98

35

29

 

 

121r

126

92

25

 

 

109

117

84

26

 

 

111

93

89

27

Test Chemical

2-Amino-4-nitrophenol

50         

69

43

141

69

71*

 

 

77

42

177

77

 

 

93

51

192

69

 

75         

114

57

251

74

73*

 

 

79

44

165

70

 

 

73

40

164

75

 

100         

92

24

206

75

82*

 

 

89

36

243

91

 

 

87

32

208

80

 

150         

93

17

365

131

118*

 

 

86

20

280

108

 

 

67

20

230

115

 

200         

86

18

287

111

133*

 

 

68

6

315

156

 

 

91

10

360

133

 

300         

100

8

393

131

143*

 

 

67

5

295

148

 

 

76

5

344

152

Positive Control

Methyl Methane Sulfonate

5         

57

46

605

357

335*

 

 

58

34

670

382

 

 

75

62

592

265

 

Footnotes: 
Asterisks(*) indicate significant responses.
r = rejected value due to quality control criteria 
# = reduced sample size because of the loss of one culture dish due to contamination 

Conclusions:
Test chemical was evaluated for its mutagenic potential in mouse lymphoma by in vitro mammalian cell gene mutation. The test result was considered to be positive in Mouse lymphoma.
Executive summary:

Genetic toxicity in vitro study was assessed for test chemical. For this purpose in vitro mammalian cell gene mutation was performed .The test material was exposed to mouse lymphoma cells.The concentration of test material used in two experiment were mention below

Experiment I; 0,25,50,100,150,200 and 300 µg/mL

 Experiment II: 50,100,150,200 and 300 µg/mL

 

Mutagenic frequency were observed in the mammalian cells. Therefore test chemical was considered to bemouse lymphoma byin vitro mammalian cell gene mutation. Hence the substance can be classified as mutagenic in vitro.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed (positive)

Genetic toxicity in vivo

Description of key information

Test substance was evaluated for its mutagenic potential in male Mouse and rat by In vivo Mammalian Somatic cell study. The test result was consider to be negative as there was no significant chromosome damage in micronucleated polychromatic erythrocytes.

Link to relevant study records
Reference
Endpoint:
in vivo mammalian somatic cell study: cytogenicity / erythrocyte micronucleus
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
data from handbook or collection of data
Justification for type of information:
data from handbook or collection of data
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)
Principles of method if other than guideline:
To evaluate the mutagenic potential of test chemical in B6C3F1 male Mouse by In vivo Mammalian Somatic cell study.
GLP compliance:
not specified
Type of assay:
other: In vivo Mammalian Somatic cell study.
Species:
mouse
Strain:
B6C3F1
Sex:
male
Details on test animals and environmental conditions:
Not specified
Route of administration:
intraperitoneal
Vehicle:
- Vehicle(s)/solvent(s) used: Corn oil
Duration of treatment / exposure:
72 hour
Frequency of treatment:
3 times in 72 hours
Remarks:
0,4.687,9.375,18.75,37.5,75,150 and 300 mg/Kg
No. of animals per sex per dose:
5 animals per sex per dose
Control animals:
yes, concurrent vehicle
Positive control(s):
Positive controls; Cyclophosphamide
- Route of administration: Intraperitoneal Injection
- Doses / concentrations:25 mg/kg
Tissues and cell types examined:
Bone Marrow cells were examined.
Evaluation criteria:
An increase in the frequency of micronucleated polychromatic erythrocytes in treated animals is an indication of induced chromosome damage.
Statistics:
Yes SD± Mean was observed.
Key result
Sex:
male
Genotoxicity:
negative
Toxicity:
not specified
Vehicle controls validity:
valid
Negative controls validity:
not specified
Positive controls validity:
valid
Remarks on result:
other: No mutagenic effect were observed

Vehicle Control

Corn Oil

 0         

5

0.700 ± 0.250

Test Chemical

2-Amino-4-nitrophenol

4.687     

5

0.900 ± 0.290

0.309

9.375     

4

1.250 ± 0.320

0.116

18.75      

5

2.100 ± 0.480

0.004

37.5       

5

1.700 ± 0.250

0.021

75         

4

1.750 ± 0.520

0.020

150         

5

1.100 ± 0.240

0.173

300         

2

1.000 ± 0.500

0.284

Positive Control

Cyclophosphamide

25         

5

16.200 ± 0.800

Value less than 0.0001

 

 

 

Dose (mg/kg)

Number of Animals Scored

Mean Percent PCE ± SEM

Pairwise P

Vehicle Control

Corn Oil

 0         

5

41.170 ± 2.800

Test Chemical

2-Amino-4-nitrophenol

4.687     

5

40.130 ± 2.570

9.375     

4

44.000 ± 1.640

18.75      

5

42.830 ± 3.470

37.5       

5

42.120 ± 2.510

75         

4

44.330 ± 2.950

150         

5

40.200 ± 3.900

300         

2

32.700 ± 8.500

Positive Control

Cyclophosphamide

25         

5

43.200 ± 2.660



 

 

Dose (mg/kg)

Number of Animals Scored

Mean MN-PCE/1000 PCE ± SEM

Pairwise P

Vehicle Control

Corn Oil

 0         

5

1.100 ± 0.190

Test Chemical

2-Amino-4-nitrophenol

18.75      

5

1.300 ± 0.250

0.342

37.5       

5

1.600 ± 0.190

0.168

75         

5

1.100 ± 0.240

0.500

150         

5

0.700 ± 0.250

0.827

Positive Control

Cyclophosphamide

25         

5

13.100 ± 0.830

Value less than 0.0001

 

 

 

Dose (mg/kg)

Number of Animals Scored

Mean Percent PCE ± SEM

Pairwise P

Vehicle Control

Corn Oil

 0         

5

46.200 ± 0.000

Test Chemical

2-Amino-4-nitrophenol

18.75      

5

54.840 ± 1.200

37.5       

5

44.600 ± 0.510

75         

5

58.640 ± 1.920

150         

5

57.320 ± 1.330

Positive Control

Cyclophosphamide

25         

5

46.960 ± 1.900

Conclusions:
Test substance was evaluated for its mutagenic potential in B6C3F1 male Mouse by In vivo Mammalian Somatic cell study. The test result was consider to be negative as there was no significant chromosome damage in micronucleated polychromatic erythrocytes.
Executive summary:

In the study test substance was assessed for its possible mutagenic potential. For this purpose wasIn vivo Mammalian Somatic cell study in B6C3F1 male Mouse. The test substance was administrated by Intraperitoneal Injection by using corn oil as vehicle. The test substance was exposed at the concentration of 0, 4.687, 9.375, 18.75, 37.5,75,150 and 300 mg/Kg for 72 hours. The dose was administrated thrice in 72 hours .The bone marrow cells were stained and observed for chromosome damage. No significant increase in the frequency of micronucleated polychromatic erythrocytes in treated animals was observed. Therefore test result was consider to be negative as there was no significant chromosome damage in micronucleated polychromatic erythrocytes. Hence the substance cannot be classified as mutant In Vivo.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (negative)

Additional information

Data for the various publication was reviewed to determine the mutagenic nature of 2-amino-4-nitrophenol (99-57-0). The studies are as mentioned below:

In Vitro studies

Ames assay

Genetic toxicity in vitro study was assessed for test chemical. For this purpose AMES test was performed .The test material was exposed to Salmonella typhimurium TA100, TA1535, TA98 and TA1537in the presence and absence of metabolic activation S9. The concentration of test material used in the presence and absence of metabolic activation were 0, 10, 30,100, 333, 1000 and 3333 µg/plate. Mutagenic effects were observed in all strains, in the presence and absence of metabolic activation. Therefore test chemical was considered to be mutagenic in Salmonella typhimurium TA100, TA1535, TA98 and TA1537by AMES test. Hence the substance can be classified as gene mutant in vitro.

Supported by other AMES test. Genetic toxicity in vitro study was assessed for test chemical. For this purpose AMES test was performed .The test material was exposed to Salmonella typhimurium TA100, TA1535, TA98 and TA1537in the presence and absence of metabolic activation S9. The concentration of test material used in the presence and absence of metabolic activation were 0, 10, 30,100, 333, 1000 and 3333 µg/plate. Mutagenic effects were observed in all strains, in the presence and absence of metabolic activation. Therefore test chemical was considered to be mutagenic in Salmonella typhimurium TA100, TA1535, TA98 and TA1537by AMES test. Hence the substance can be classified as gene mutant in vitro.

 

In vitro chromosomal abbreviation study

Genetic toxicity in vitro study was assessed for test chemical. For this purpose in vitro mammalian chromosome aberration test was performed .The test material was exposed toChinese hamster ovary cells inthe presence and absence of metabolic activation S9. The concentration of test material used in the presence and absence of metabolic activation were mention below

-S9;0,150,199,249and 300 µg/mL

+S9; 0,2460,2760,2990 and 3520 µg/mL

Chromosome aberration, Chromosome gaps and breaks were observed in the presence or absence of metabolic activation. Therefore test chemical was considered to be mutagenic inChinese hamster ovary cells byin vitro mammalian chromosome aberration test. Hence the substance cannot be classified as mutagenic in vitro.

In vitro Mammalian cell gene mutation assay

Genetic toxicity in vitro study was assessed for test chemical. For this purpose in vitro mammalian cell gene mutation was performed .The test material was exposed to mouse lymphoma cells.The concentration of test material used in two experiment were mention below

Experiment I; 0,25,50,100,150,200 and 300 µg/mL

 Experiment II: 50,100,150,200 and 300 µg/mL

 

Mutagenic frequency were observed in the mammalian cells. Therefore test chemical was considered to be mutagenic in mouse lymphoma by in vitro mammalian cell gene mutation. Hence the substance can be classified as mutagenic in vitro.

In vitro DNA damage and/or repair study

Genetic toxicity in vitro study was assessed for test chemical. For this purpose in vitro sister chromatid exchange assay was performed .The test material was exposed toChinese hamster ovary cells inthe presence and absence of metabolic activation S9. The concentration of test material used in the presence and absence of metabolic activation were mention below

-S9;0,5,16.7 and 50 µg/mL

+S9; 0,1740,2180 and 2670 µg/mL

 

The mammalian cells were observed fornumber of scored exchanges correlates to the number of DNA breakage and reunion eventsin the presence or absence of metabolic activation. The test article induced sister chromatid exchange in the cultured cells.  Therefore test chemical was considered to be mutagenic inChinese hamster ovary cells byin vitro sister chromatid exchange assay. Hence the substance cannot be classified as mutagenic in vitro.

In Vivo studies

In the study test substance was assessed for its possible mutagenic potential. For this purpose wasIn vivo Mammalian Somatic cell study in B6C3F1 male Mouse. The test substance was administrated by Intraperitoneal Injection by using corn oil as vehicle. The test substance was exposed at the concentration of 0, 4.687, 9.375, 18.75, 37.5,75,150 and 300 mg/Kg for 72 hours. The dose was administrated thrice in 72 hours .The bone marrow cells were stained and observed for chromosome damage. No significant increase in the frequency of micronucleated polychromatic erythrocytes in treated animals was observed. Therefore test result was consider to be negative as there was no significant chromosome damage in micronucleated polychromatic erythrocytes. Hence the substance cannot be classified as mutant In Vivo.

In the study test substance was assessed for its possible mutagenic potential. For this purpose wasIn vivo Mammalian Somatic cell study in Fischer 344 male rat. The test substance was administrated by Intraperitoneal Injection by using corn oil as vehicle. The test substance was exposed at the concentration of 0, 3.91, 7.815, 15.63, 31.25, 62.5 and 125 mg/Kg for 72 hours. The dose was administrated thrice in 72 hours .The bone marrow cells were stained and observed for chromosome damage. No significant increase in the frequency of micronucleated polychromatic erythrocytes in treated animals was observed. Therefore test result was consider to be negative as there was no significant chromosome damage in micronucleated polychromatic erythrocytes. Hence the substance cannot be classified as mutant In Vivo.

 

 

 

Based on the data summarized, 2-amino-4-nitrophenol (99-57-0)did  not induce gene mutation in vivo while it induce gene mutation in vitro.The test chemical induce mutation in AMES test as well as in OECD 473 test (In vitro chromosomal abbreviation study) . The test chemical induce gene mutation in In vitro Mammalian cell gene mutation assay too. But as the test chemical did not induce gene mutatiomn in IN vivo study conducted in male rat and male mouse.As per CLP criteria

The classification in Category 2 is based on:

— positive evidence obtained from experiments in mammals and/or in some cases from in vitro experiments, obtained from:

— somatic cell mutagenicity tests in vivo, in mammals; or

— other in vivo somatic cell genotoxicity tests which are supported by positive results from in vitro mutagenicity assays.

Hence based on CLP criteria the test substance can not be classified as mutagenic.

Justification for classification or non-classification

Based on the data summarized and CLP criteria , 2-amino-4-nitrophenol (99-57-0) not likly induce gene mutation in vivo while it induce gene mutation in vitro.The test chemical induce mutation in AMES test as well as in OECD 473 test (In vitro chromosomal abbreviation study) .

The test chemical induce gene mutation in In vitro Mammalian cell gene mutation assay too.

But as the test chemical did not induce gene mutatiomn in In vivo study conducted in male rat and male mouse.As per CLP criteria

The classification in Category 2 is based on:

— positive evidence obtained from experiments in mammals and/or in some cases from in vitro experiments, obtained from:

— somatic cell mutagenicity tests in vivo, in mammals; or

— other in vivo somatic cell genotoxicity tests which are supported by positive results from in vitro mutagenicity assays.

As no evidence of somatic cell mutagenicity tests in vivo, in mammalswas observed

Hence based on CLP criteria the test substance not likly to be be classified as mutagen.